NFLX抗原模拟表位的淘选及鉴定  

Biopanning and identification of norfloxacin mimotope

作　　者：熊争平[1] 许杨[1,2] 何庆华[2] 

机构地区：[1]南昌大学食品科学与技术国家重点实验室,江西南昌330047 [2]南昌大学中德联合研究院,江西南昌330047

出　　处：《南昌大学学报（理科版）》2013年第4期377-381,共5页Journal of Nanchang University(Natural Science)

基　　金：国家自然科学基金项目(31171696;31201360);广东省教育部产学研结合项目(2010B09040182)

摘　　要：从噬菌体随机肽库中淘选模拟NFLX（Norfloxacin）表位的噬菌体粒子并进行性质鉴定。以抗NFLX单克隆抗体为靶分子，对噬菌体环七肽库和十二肽库进行生物亲和淘选，以ELISA鉴定阳性克隆，进行DNA测序，采用生物信息学方法对淘选得到的模拟表位进行分析。分别得到了9个（环七肽）和8个（十二肽）能与靶分子特异性结合且能被NFLX标准品阻断的阳性克隆，其特有序列为（环七肽）：X-X-X-X-X-脯氨酸-苯丙氨酸（X为任意氨基酸），以阻断效果最好的抗原模拟表位H3建立了竞争ELISA标准曲线，IC50为：（48．07±6）ng·mL^-1，检测线性范围为10-200ng·mL^-1。噬菌体展示技术可淘选得到NFLX模拟表位，淘选到的噬菌体粒子可作为NFLX的替代品建立免疫学检测方法。To screen phage particles capable of mimicking norfloxacin （NFLX） and identify the properties. The anti-NFLX monoclonal antibody was used as a target for biopanning from c7c phage display peptide library and phage 12-mer peptide library. The positive clones were identified by ELISA and sequenced, then analyse the obtained mimotopes by means of the bioinformatics. 9 positive clones（c7c phage display peptide library）and 8 positive clones（phage 12-mer peptide library）can bind with anti-NFLX monoclonal antibody and be blocked by NFLX,they have common feature（c7c phage display peptide）. X-X-X-X-X-prolin-phenylalanine （X can be any amino acid）, H3 was used to establish a competitive ELISA immunoassay,the linear range of the inhibition curves was between 10 ng . mL^-1 and 200 ng. mL^-1 ,IC50 was 48.07±6 ng. mL^-1. The phage display technique can be successfully applied to panning the mimotope of Norfloxacin. The aquired phages may be used as the surrogate of the toxin to establish immunoassay.

关 键 词：噬菌体随机肽库 NFLX 模拟表位 免疫学检测方法 

分 类 号：Q782[生物学—分子生物学]